PCR–ELISA for High-Throughput Blood Group Genotyping

Abstract

During the last decade, blood bank specialists have shown an increased interest in molecular analyses to complement serology work in determining blood group antigens. To efficiently respond to the numerous demands made for hemolytic disease of the newborn cases and polytransfused patients, we designed an inexpensive colorimetric high-throughput method to genotype several blood group antigens rapidly. Three simple steps are required to perform this technique: genomic DNA extraction, PCR amplification, and amplicon detection by a microplate ELISA. The 96-well plate format facilitates the manipulations and enables the analysis of multiple samples at once or the analysis of multiple antigens for fewer samples.

The most common and clinically relevant minor blood group antigens were adapted to this method and are described in this work: Rh (D, C, c, E, e), Kell (K, k), Duffy (Fya, Fyb), and Kidd (Jka, Jkb). Other blood group antigens could be easily tested this way as long as their molecular basis is well established.